1475

Journal of Food Protection, Vol. 76, No. 8, 2013, Pages 1475–1479

doi:10.4315/0362-028X.JFP-12-561
Copyright G, International Association for Food Protection

Research Note

Evaluation of Microbial Contamination of Tomatoes and Peppers

at Retail Markets in Monterrey, Mexico
CARMEN CÁRDENAS, KARINA MOLINA, NORMA HEREDIA, AND SANTOS GARCÍA*

Departamento de Microbiologı́a e Inmunologı́a, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Avenida Universidad s/n,
Ciudad Universitaria, San Nicolás de los Garza, Nuevo León, CP 66451, Mexico

MS 12-561: Received 18 December 2012/Accepted 7 March 2013

ABSTRACT
The source of a large outbreak of foodborne disease related to Salmonella-contaminated jalapeño peppers has been traced to
Nuevo Leon, Mexico. The objective of this work was to evaluate the microbiological quality of tomatoes and jalapeño peppers
from markets and supermarkets from the metropolitan area of Monterrey, Nuevo León, Mexico. One hundred sixty samples (40
bola tomatoes, 40 saladette [Roma] tomatoes, 40 serrano peppers, and 40 jalapeño peppers) were purchased. Stems from peppers
were removed and analyzed separately. Samples were analyzed for indicator organisms and Salmonella, following the Mexican
Official Methods. The results showed that the presence of indicator organisms varied among samples and origins, and levels were
relatively high in peppers (average 4.4 to 4.7 log CFU/g for total mesophilic, 3.25 to 3.73 log CFU/g for total coliforms, and
1.69 log CFU/g for fecal coliforms). Saladette tomatoes and serrano peppers showed the greatest microorganism levels
(,1 log CFU/g higher) in comparison with the other varieties. Pepper stems typically had indicator microbial levels ,1 to
2 log CFU/g higher than levels in smooth flesh. Only one tomato and one jalapeño sample were positive for Salmonella.
However, in the case of the pepper, the contamination was found in the stem. Although the microbiological quality of tomatoes
and peppers sampled was similar to that found in markets from developed countries, the presence of pathogens causes a risk of
infection for consumers.

Consumption of fresh produce has increased within the
past decade, mainly because of awareness of the benefits of
a healthy diet and the impact of governmental campaigns in
several countries, such as the ‘‘Five a Day’’ and ‘‘Nine a
Day’’ programs in the United Kingdom and the United
States, and global efforts, such as those by the World Health
Organization (29). However, fresh vegetables can be
contaminated with pathogens from animal and human
sources during growth, harvest, transportation, and further
handling (11, 20). Contaminated soil, animal fecal drop-
pings, poor worker hygiene, inadequately composted or raw
animal manure or sewage, poor water quality for irrigation
or washing, etc., contribute to the possible contamination of
produce with microbial pathogens; in general, lapses in
safety practices can produce product contamination (3, 4,
13, 20, 24).
Due to these contributing factors, produce has been
involved in an increasing number of foodborne disease
outbreaks (18). This increase in outbreaks can also be
attributed to changing consumer habits, new methodologies
to detect microorganisms, improved surveillance, and the
globalization of the produce trade (20).
The epidemiology of foodborne disease has changed
rapidly over the last decades; and some major human
* Author for correspondence. Tel/Fax: z52-818-3763044; E-mail:
santos@microbiosymas.com.
pathogens, such as Escherichia coli O157:H7, Salmonella,
and Campylobacter, have been recognized to be spread
from animal reservoirs (15). Fresh vegetables have emerged
as vehicles for the transmission of these infectious diseases
(11). Leafy greens, tomatoes, cucurbits, peppers, and nuts
have been among the foods commonly linked to outbreaks
of gastrointestinal illnesses caused by E. coli O157:H7 and
Salmonella (17). Salmonella spp. are the most commonly
identified etiological agents associated with fresh produce–
related infection (7). At least 12 well-documented outbreaks
in the United States have involved tomatoes; tomatoes were
responsible for 1,990 confirmed cases and were suspected in
another 75,000 cases (7, 19, 27). Outbreaks have involved
diverse Salmonella serotypes, including Javiana, Montevi-
deo, Baildon (16), Typhimurium (6), and Wandsworth (7).
In 2008, a multistate outbreak in the United States and
Canada was caused by Salmonella Saintpaul. Jalapeño
peppers were implicated in this outbreak, but serrano
peppers and tomatoes also were believed to have contrib-
uted to it. The outbreak strain was detected in jalapeño
peppers collected in Texas and also in water and serrano
peppers on a Mexican farm in the state of Nuevo Leon.
Tomato tracebacks did not converge on a source (2).
Chili peppers are important commercial crops in
Mexico; in 2009, more than 2 million tons were produced.
Of this production, approximately 25 and 10% were
jalapeño and serrano peppers, respectively (26). These two
1476 CÁRDENAS ET AL.
pepper species are most commonly consumed raw in
Mexico and other countries (10). Tomato production in
Mexico increased by 56.2% from 1994 to 2001, and during
the winter, most of the tomatoes consumed in the United
States come from Mexico (18).
Produce imported from Mexico, although it has been
shown to be of an equivalent microbial quality to domestic
samples in the United States (18), has been linked to several
disease outbreaks in the United States (5, 8, 9). Intestinal
diseases due to consumption of contaminated food are
among the top five causes of disease in Mexico (9). Studies
in central Mexico have found high levels of contamination
on serrano and jalapeño peppers (10, 25). Another study in
northeastern Mexico conducted a brief analysis of different
crops, including tomatoes and peppers, and found no
pathogens (14). However, that study did not analyze
different varieties of tomatoes. The current study investi-
gates the microbiological quality of two varieties of
tomatoes (saladette and bola) and two varieties of peppers
(jalapeño and serrano) in several retail establishments in the
metropolitan area of Monterrey, in the state of Nuevo Leon,
Mexico, during 2010 to 2011. Of particular interest was the
analysis of pepper stems, which have been reported to have
high levels of contamination (21).
MATERIALS AND METHODS
Vegetable samples. The microbial contamination level of 160
produce samples of four products (serrano peppers, jalapeño
peppers, bola tomatoes, and saladette [Roma] tomatoes) was
assessed. Forty samples of each item were obtained (20 from local
markets and 20 from supermarkets) from October 2010 to February
2011. Samples were collected in the metropolitan area of
Monterrey, Mexico (including the cities of Monterrey, Guadalupe,
San Nicolás, Escobedo, and San Pedro), placed in sterile bags, and
transported on ice within 1 h to the laboratory. Samples were
analyzed immediately upon arrival.
Sample preparation and microbial determination. Upon
arrival at the laboratory, stems were separated from the peppers,
and samples were analyzed separately. Samples (25 g) from each
produce item were randomly removed and homogenized with
225 ml of buffered peptone water and then were pummeled for
2 min in a stomacher lab blender (Led Techno, Eksel, Belgium);
serial dilutions were made from the blended material. For the
pepper stems, 10 g of sample was homogenized with 90 ml of
buffered peptone water, and the same protocol was followed.
To determine the level of aerobic mesophilic microorganisms,
the Mexican official standard protocol (NOM-092-SSA1-1994
(22)) was used. Briefly, 1 ml of each serially diluted sample was
placed into a petri dish. Then 20 ml of plate count agar (Difco, BD,
Sparks, MD) heated to 45uC was added, and the mixture was
gently homogenized. After solidification, plates were incubated
(35 ¡ 1uC for 48 h), and colonies were counted.
For coliform and E. coli determination, the method described
in the Bacteriological Analytical Manual (12) was followed. The
serially diluted sample (1 ml) was placed in the center of a petri
dish, 20 ml of violet red bile agar (Difco, BD, heated at 45uC) was
added, and the mixture was gently homogenized. After solidifica-
tion, the agar was overlaid with 5 ml of double-strength violet
red bile agar supplemented with 4-methylumbelliferyl-b-D-
glucuronide and was allowed to solidify. Inverted plates were
J. Food Prot., Vol. 76, No. 8
incubated at 35uC for 24 ¡ 2 h, and red-purple colonies were
counted and considered as total coliforms, whereas fluorescent
colonies under UV light were considered to be E. coli. To
determine fecal coliforms, red-purple colonies were inoculated in
tubes containing 5 ml of E. coli broth (BD Diagnostic Systems,
Franklin Lakes, NJ) with a Durham tube. After 48 h at 45uC, tubes
showing growth and gas production were considered as positive.
To determine the presence of Salmonella, the Mexican official
standard protocol (NOM-114-SSA1-1994 (23)) was used. For
preenrichment, samples (25 g) were transferred into a sterile
Whirl-Pak bag (Nasco, Fort Atkinson, WI), and 225 ml of sterile
lactose broth was added. For pepper stem analysis, 10 g of stems and
90 ml of lactose broth were used. After mixing well by swirling, the
pH was adjusted to 6.8 ¡ 0.2, followed by incubation for 24 ¡ 2 h
at 35uC. Enrichment was made by transferring 1 ml of the mixture
into a tube with 10 ml of Rappaport-Vassiliadis medium and 10 ml
of selenite-cystine broth (both from Difco, BD). Incubation was
carried out at 35 ¡ 0.2uC for 24 h. Next, a loopful of culture was
streaked onto plates of xylose lysine desoxycholate agar, Salmonella
Shigella agar, and brilliant green agar (all from Difco, BD). Plates
were incubated for 24 ¡ 2 h at 35uC. Colonies suggestive of
Salmonella on xylose lysine desoxycholate agar (pink colonies with
or without black centers), brilliant green agar (pink to fuchsia
colonies surrounded by red medium), or Salmonella Shigella agar
(transparent to opaque colonies with or without black centers) were
harvested and subjected to additional tests (lactose, glucose, and
sucrose fermentation, lysine decarboxylation, sulfhydric acid, and
urease production) and screened for the presence of the invA gene by
PCR (28)). Salmonella Typhi ATCC 19430 and Salmonella
Typhimurium ATCC 14028 were used as positive controls.
Statistical analysis. To determine whether the samples were
normally distributed, the Kolmogorov-Smirnov test was used (30).
For mesophilic and total coliform microorganisms, one-way
analysis of variance (30) was conducted to determine differences
in levels among produce types, location, and kind of market. For
fecal coliforms, the Kruskal-Wallis test (30) was followed, and for
Salmonella spp., the Cochran test (30) was used. Differences were
considered to be significant if P # 0.05. Analyses were performed
using SPSS Statistics software (version 17.0, SPSS, Inc., Chicago,
IL) and STATISTICA (version 7.0, Statsoft, Tulsa, OK).
RESULTS AND DISCUSSION
Detection of pathogenic microorganisms on contami-
nated produce is difficult for a variety of reasons, including
the long time required, complicated methodologies, and
high cost. The detection of representative indicator
microorganisms is easier and may be used to signal the
presence of pathogenic bacteria (21). Our results showed
that total viable counts (aerobic mesophilic microorganisms)
for crops ranged from fewer than 10 to greater than
105 CFU/g (Figs. 1 to 3). Levels were higher in peppers
(means of 4.4 and 4.7 log CFU/g for jalapeño and serrano,
respectively) than in tomatoes; both varieties of tomato
showed low levels of bacteria (means of 3.2 and
3.6 log CFU/g for bola and saladette, respectively)
(Fig. 4). Differences (P # 0.05) in microbial load among
produce varieties of both vegetables were observed; serrano
peppers and saladette tomatoes had the highest levels.
However, no differences were found between the market
types (data not shown). The results obtained are in
accordance with those reported by Gómez et al. (14) for
J. Food Prot., Vol. 76, No. 8 MICROBIOLOGICAL QUALITY OF TOMATOES AND PEPPERS FROM MEXICO
FIGURE 1. Total viable counts of microorganisms (TVC), total
coliforms (TC), and fecal coliforms (FC) present in jalapeño
peppers and jalapeño stems, purchased in the metropolitan area of
Monterrey, Mexico. Upper number indicates the number of
positive samples.
vegetables in northeastern Mexico; they reported counts
lower than 104 CFU/g on tomatoes and reported differences
in levels of mesophilic aerobic bacteria among different
vegetables. Similarly, Liao et al. (21) reported that peppers
obtained from grocers in Pennsylvania had levels of aerobic
microorganisms ranging from 4.7 to 6.3 log CFU/g of
tissue, with an average of 5.6; these levels are similar to
those found in the present study. Another study conducted
in the central part of Mexico (Pachuca, Hidalgo) reported
higher levels of mesophilic aerobic bacteria in peppers at
retail with counts of 7.2 and 6.4 log CFU/g for serrano and
jalapeño peppers, respectively (10). This level of contam-
ination could be due to deficiencies in quality control
systems of pepper production and distribution in that region.
The total coliform levels found in this study for all
crops ranged from less than 10 to greater than 105 CFU/g
(Figs. 1 to 3), with median values of 3.3, 3.7, 2.6, and
2.6 log CFU/g for jalapeño, serrano, bola, and saladette
varieties, respectively (Fig. 5). Peppers (jalapeño and
serrano) exhibited greater concentrations of coliform
bacteria than tomatoes (Fig. 5); however, no differences
(P # 0.05) were observed among varieties of each crop or
the type of store where the samples were purchased. The
levels of contamination found in this work were much less
than those reported for jalapeño (3.0 to 8.1 log CFU per
sample) and serrano peppers (log 4.5 to 8.5 CFU per
sample) from popular markets in Pachuca, Hidalgo, Mexico
1477
FIGURE 2. Total viable counts of microorganisms (TVC), total
coliforms (TC), and fecal coliforms (FC) present in serrano
peppers and serrano stems, purchased in the metropolitan area of
Monterrey, Mexico. Upper number indicates the number of
positive samples.
(10). On the other hand, the levels of fecal coliforms ranged
from less than 10 to 103 log CFU/g for peppers and less than
10 to 102 log CFU/g for tomatoes (Figs. 1 to 3). In addition,
the levels of fecal coliforms in this study were slightly less
than those reported for jalapeño and serrano peppers in the
central part of Mexico (11). These differences in contam-
ination could be due to variations in agricultural and
handling practices as well as analysis methods.
Pepper stems accounted for a greater contamination
level of aerobic mesophilic and total coliforms, when
compared to the rest of the fruit (Figs. 1 and 2). These
results are in accordance with those reported by Liao et al.
(21), who determined the relative distribution of native
microflora on two different parts of jalapeño peppers (calyx
[stem] and flesh pod) and demonstrated that most of the
contamination sites were located at the calix (stem)
(.80%), while only a small proportion (,10%) was
recovered from the flesh pod. The wrinkled texture of the
stem may better suit it to harbor microorganisms than the
smooth texture of the comestible area. Furthermore, the
stem is the contact area of pepper used to pluck it from the
plant during harvest.
Mexico is divided into three major regions for pepper
production: the north-northeast region with high levels of
technology, the central (Bajı́o) region with medium levels of
1478 CÁRDENAS ET AL.
FIGURE 3. Total viable counts of microorganisms (TVC), total
coliforms (TC), and fecal coliforms (FC) present in bola and
saladette tomatoes purchased in the metropolitan area of
Monterrey, Mexico. Upper number indicates the number of
positive samples.
technology, and the south-southeast region with low levels
of technology (1). These conditions could also help to
explain the differences in contamination of peppers found in
central and northern Mexico.
FIGURE 4. Total viable counts of microorganisms present in
fresh produce purchased in the metropolitan area of Monterrey,
Mexico. The box represents 75% of the samples. The line inside
each box indicates the geometric mean.
J. Food Prot., Vol. 76, No. 8
FIGURE 5. Total coliform counts present in fresh produce
purchased in the metropolitan area of Monterrey, Mexico. The
box represents 75% of the samples. The line inside each box
indicates the geometric mean.
Historically, outbreaks of salmonellosis in humans have
been attributed to consumption of contaminated tomatoes,
mustard cress, bean sprouts, cantaloupe, and watermelon.
However, in the summer of 2008, consumption of
Salmonella-contaminated jalapeño peppers was implicated
in one of the largest reported foodborne illness outbreaks
(8). Analysis of the presence of pathogenic bacteria in our
samples showed that only one sample of bola tomatoes
(1.25%) and one of jalapeños (1.25%) were positive for
Salmonella (confirmed by PCR). However, the pepper
contamination was found in the stem, which usually is
removed before consumption. The frequency of positive
samples for Salmonella in tomatoes and peppers is usually
low (19), which is in accordance with our results. In
conclusion, the results of our study show that the level of
contamination of peppers and tomatoes is low in the
metropolitan area of Monterrey, Mexico.
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